Nuclear and Mitochondrial Circulating Cell-Free DNA Is Increased in Patients With Inflammatory Bowel Disease in Clinical Remission

Zuzana Vrablicova, Kristina Tomova, Lubomira Tothova, Janka Babickova, Barbora Gromova, Barbora Konecna, Robert Liptak, Tibor Hlavaty, Roman Gardlik, Zuzana Vrablicova, Kristina Tomova, Lubomira Tothova, Janka Babickova, Barbora Gromova, Barbora Konecna, Robert Liptak, Tibor Hlavaty, Roman Gardlik

Abstract

Background: The role of cell-free DNA (cfDNA) in the pathogenesis of inflammatory bowel disease (IBD) has been recently suggested. The aim of this study was to analyze circulating cfDNA and deoxyribonuclease (DNase) activity in IBD patients in clinical remission. Materials and Methods: Plasma and serum were obtained from 72 patients with Crohn's disease and 28 patients with ulcerative colitis. Total cfDNA, nuclear DNA (ncDNA), mitochondrial DNA (mtDNA) and DNase activity were measured. Results: IBD patients showed higher levels of both ncDNA and mtDNA compared to healthy controls. Concentration of ncDNA was higher in males compared to females, including patients and healthy controls. However, unlike males higher amount of ncDNA was found in female IBD patients compared to healthy controls. DNase activity was significantly lower in male IBD patients compared with healthy controls. In addition, there was a negative correlation between DNase activity and ncDNA levels in male IBD patients. Conclusions: Herein we present increased amount of circulating ncDNA and mtDNA in IBD patients in clinical remission. Thus, unlike total cfDNA, circulating ncDNA and mtDNA might not represent the optimal biomarkers of disease activity. This is also the first report on sex difference in circulating ncDNA levels, possibly associated with lower DNase activity in males.

Keywords: Crohn's disease; deoxyribonuclease (DNase); extracellular DNA; remission; ulcerative colitis.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2020 Vrablicova, Tomova, Tothova, Babickova, Gromova, Konecna, Liptak, Hlavaty and Gardlik.

Figures

Figure 1
Figure 1
Concentration of total cfDNA, ncDNA and mtDNA in IBD patients and controls. No difference in total cfDNA between healthy controls and patients was found (A,B). IBD patients showed higher levels of both ncDNA and mtDNA compared to healthy controls (C: p = 0.0061 and E: p = 0.0387). The difference was also seen in both subgroups of CD and UC patients (D: p = 0.0062 for CD and 0.0105 for UC against control and F: p = 0.0345 for CD and p = 0.0365 for UC against control). *P < 0.05, **P < 0.01.
Figure 2
Figure 2
Sex differences in cfDNA, ncDNA and mtDNA. Higher total cfDNA was found in male IBD patients compared with female patients (p = 0.0478); however, this difference was not found in the subgroups of CD and UC (A). Concentration of ncDNA is higher in males compared to females in healthy controls (p = 0.0274), IBD patients (p = 0.0345) and CD subgroup (p = 0.0309) (B). No sex difference was found in mtDNA in any of the groups (C). *P < 0.05.
Figure 3
Figure 3
Total cfDNA, ncDNA and mtDNA in females. No difference was found between groups in total cfDNA (A). Higher amount of ncDNA was found in female IBD patients (p = 0.0098), including both CD (p = 0.0108) and UC (p = 0.0110) compared to healthy controls (B). Similar trend is seen in mtDNA, although the differences were not significant (C). *P < 0.05, **P < 0.01.
Figure 4
Figure 4
Total cfDNA, ncDNA and mtDNA in males. No significant differences between groups were shown in cfDNA (A), although a clear trend toward higher ncDNA and mtDNA in IBD patients compared with healthy controls can be seen (B,C).
Figure 5
Figure 5
DNase activity in healthy controls and patients. In females, no difference was found between groups. DNase activity was lower in male IBD patients (p = 0.0001), including both CD (p = 0.0002) and UC (p = 0.0497) compared with healthy controls (A). Negative correlation was found between DNase activity and concentration of ncDNA in male IBD patients (p = 0.0198) (B). *P < 0.05, ***P < 0.001.

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